Molecular and Structural Insights into Buffalo Interleukin-17A.

Interleukin-17A is a pro-inflammatory cytokine that plays a key role in the immune response to many pathogens and implicated in autoimmune diseases. This molecule is also involved in providing protection to many bacterial and fungal infections of gastro-intestinal tract and respiratory mucosa. Although molecular aspect of IL-17A has been studied in few species, no data are available for buffalo, which is one of the major sources of milk production in India. Therefore, in the present study, IL-17A gene of Indian Murrah Buffalo origin was cloned, expressed, and analyzed using bioinformatic tools. The coding sequence of buffalo IL-17A gene was cloned in prokaryotic expression vector (pET-28a) followed by its expression, purification, and characterization. A computational analysis was performed to understand the sequence, structure, and evolutionary relationship of buIL-17A. It revealed that the length of buIL-17A sequence without signal peptide is 132 amino acids as in cattle. However, sequence identity is found to be 99% due to one amino substitution difference between buffalo and cattle. After analysis, it can be concluded that buIL-17A recombinant protein can be used as a potential immunobiological reagent for diagnostic and therapeutic purpose.

Characterization of Foot-and-Mouth Disease Virus Serotype O-Specific Single Domain Antibody Expressed in the pET Expression System.

Foot and mouth disease (FMD) is an extremely contagious disease of cloven-hoofed domesticated and wild animals, resulting in significant economic losses in many parts of the world. FMD virus (FMDV) serotype O is responsible for approximately 70% of global outbreaks. For detection of FMDV antigen or antibody, ELISAs are used worldwide and have several limitations, such as batch-to-batch variation in generating immunobiologicals, high production cost and ethical concerns over animal sacrifice. The use of single domain antibody (sdAb) or variable N-terminal domain of the heavy chain of heavy chain antibody (VHH) found naturally in camels has proven their effectiveness in diagnostics and therapeutics. In the present study, the anti-FMDV serotype O-specific VHH-C1 gene sequence (Accession no. KJ751546) was retrieved from the NCBI database. The gene was synthesized commercially in the pBluescript KS+ cloning vector and expressed in E. coli BL21 (DE3) cells using the pET303/CT-His expression system with a C-terminal 6X-His tag. The expressed sdAb, verified by SDS‒PAGE and western blotting, was purified by Ni-chelate chromatography and used as a coating antibody in double antibody sandwich (DAS) ELISA for FMDV detection and typing. The sdAb exhibited a high binding affinity for FMDV serotype O, without any cross-reactivity toward serotypes A and Asia-1. It exhibited better thermostability up to 85 °C than conventional rabbit polyclonal anti-FMDV sera. The potential of sdAbs thus produced without sacrificing lab animals could be explored for replacing polyclonal sera in DAS-ELISA as well as for developing biosensors or lateral flow devices for FMDV type O detection.

Clinical Diagnosis and Therapeutic Management of Trypanosoma theileri Infection Associated with Peritonitis in Crossbred Cow: A Rare Case Report.

PURPOSE: The present study reports a very rare case of Trypanosoma theileri like infection in the peritoneal fluid of a 6-year-old female Holstein Friesian (HF) cattle brought to Veterinary Clinical Complex, Lala Lajpat Rai University of Veterinary and Animal Sciences (LUVAS), Hisar. The aim of the clinical study was to report an unusual case of T. theileri-like infection associated with peritonitis and its therapeutic management. METHODS: For confirmatory diagnosis, peritoneal fluid and blood sample was collected from animal for haemato-biochemical examination, buffy coat examination and detection of Trypanosoma antigen by monoclonal antibody-based latex agglutination test. RESULTS: Animal had history of anorexia, high fever, oedematous swelling on the abdominal area and decrease in milk yield since 1 week. Clinical examination revealed high fever, congested conjunctival mucus membrane and oedematous inflammatory swelling on the abdominal region. Haematological examination on Day 1 revealed relative neutrophilia and eosinophilia, whereas other parameters measured were on the lower side of normal range. Thin blood smear examination did not reveal any parasite on first attempt. However, after confirmation of Trypanosoma species in peritoneal fluid collected by FNAC, buffy coat examination of blood sample was also found positive for Trypanosoma species. On Day 2, haematological examination revealed neutrophilic leucocytosis, eosinophilia and anaemia. Blood sample was also found serologically positive for Trypanosoma antigen by monoclonal antibody based latex agglutination test. Biochemical examination revealed increased blood urea nitrogen (BUN), gamma glutamyl transferase (GGT) levels, hyperglycemia, hyperprotienemia and decreased levels of sodium and chloride ions. Animal was primarily treated with isometamidium chloride (Nyzom®) @ 0.5 mg/kg body weight (IM, single dose) along with supportive therapy. CONCLUSION: Animal showed clinical recovery after 10 days of treatment. Post-recovery, blood sample of the animal was re-examined for parasite and other blood parameters and it was found negative for presence of Trypanosoma infection along with restoration of haematological values. Hence, the present study concludes the confirmatory diagnosis and therapeutic use of isometamidium hydrochloride along with anti-inflammatory drugs to Trypanosoma infection.

Molecular detection and characterization reveals circulation of multiple genotypes of porcine astrovirus in Haryana, India.

Porcine astrovirus (PAstV) is distributed worldwide and has been reported to cause diarrhea in pigs. PAstV belongs to the family Astroviridae and genus Mamastrovirus. PAstVs are divided into five diverse genotypes (PAstV1-PAstV5) on the basis of phylogenetic analysis of a part of the RNA-dependent RNA polymerase (RdRp) gene and the capsid gene. However, knowledge regarding the clinical significance and molecular characteristics of PAstV in Haryana, India, is limited. In this study, we investigated the presence of PAstV by RT-PCR of the partial RdRp gene in 110 rectal swabs collected from diarrheic pigs in different parts of Haryana, India. Of these, 35 samples (31.8%) tested positive for PAstV, with the highest positivity observed among weaning piglets 3 to 9 weeks of age (47.7%, 21/44), followed by fattening pigs 9 to 24 weeks of age (28.5%, 8/28). Phylogenetic analysis of the partial RdRp gene revealed circulation of four different genotypes (PAstV1, PAstV2, PAstV4, and PAstV5) in Haryana, with PAstV1 being the predominant genotype. To the best of our knowledge, this is the first report of the presence of PAstV1 and PAstV5 in the pig population of India. The PAstV sequences revealed high genetic variability and genetic heterogeneity in a relatively confined area.

Digital gangrene induced by inadvertent intra-arterial cocktail injection of anesthetic agents such as pentazocine, promethazine, and atropine: A serious adverse drug experience.

Gangrenous changes in skin due to accidental intra-arterial injection of promethazine and pentazocine have been reported. Accidental intra-arterial injection is most commonly encountered in the antecubital fossa. However, recent reports in the radial and ulnar arteries have also been encountered. We hereby report a serious, preventable adverse drug experience in the form of digital gangrene induced by inadvertent intra-arterial cocktail injection of anesthetic agents such as pentazocine, promethazine, and atropine, which seems to be in the radial artery as the lateral three digits and dorsum of the hand are affected.

Full-Genome Sequence Analysis of a Reassortant Strain of Bluetongue virus Serotype 16 from Southern India.

The complete genome sequence of a reassortant field strain (IND2014/01) of Bluetongue virus (BTV) serotype 16, isolated from sheep from southern India in 2014, was sequenced. The total genome size was 19,186 bp. Sequence comparisons of all genome segments, except segment 5 (Seg-5), showed that IND2014/01 belonged to the major eastern topotype of BTV.